IS 12572 ( Part 6 ) : 1988

Indian Standard

( Reaffirmed 2006 )

GUIDE FOREVALUATIONOFMEDICAL DEVICES FOR BIOLOGICAL HAZARDS
PART 6 METHOD OF TEST FOR SYSTEMIC TOXICITY : ASSESSMENT OF PYROGENICITY IN RABBITS OF EXTRACTS FROM MEDICAL DEVICES

UDC

615'4 : 614'8 : 616-02~615'009~092

DESCRIPTORS : MEDICAL DEVICES. BIOLOGICAL HAZARDS. SYSTEMIC TOXICITY. PYROGENICITY.

Q BIS 1989

BUREAU
MANAK

OF

INDIAN

STANDARDS

H H A V A N , 9 BAHADUt SllAH ZAFAR MARG NEW DELHI I IWO2 Price Group 2

Sqmnber 1989

Medical Glass Instruments and Appliances Seclional Committee, CPDC 12

FOREWORD This Indian Standard ( Part 6 ) was adopted by the Bureau of Tndian Standards on 14 September 1988, after the draft finalized by the Medical Glass Instruments and Appliances Sectional Committee had been approved by the Consumer Products and Medical Instruments Division Council. This part ( Part 6 ) of the standard is one of a series of methods of test for assessing the biological hazards of medical devices that directly contact body tissues or substances that are to be introduced into the body by means other than oral. The test in this standard has been selected to indicate the presence of pyrogenic substances in or on a device, although it is recognized that pyrogens of microbial origin will be detected, if present. In some cases, the pyrogenicity caused by gram negative endotoxaemia can be detected by the LAL test which can be employed as an additional test. This, however, does not exclude some of the requirements of the pyrogen test, particularly when LAL test is negative. This standard is proposed to be published in 12 parts. Other parts are as follows: Part 1 Part 2 Part 3 Part 4 Part 5 Part 7 Part 8 Part 9 Part 10 Part 11 Part 12 Terminology Selection of biological methods of test Method of testing by tissue implantation Method of test for systemic toxicity: Assessment of acute toxicity of extracts from medical devices Method of test for intracutaneous reactivity of extracts from medical devices Methods of test for sensitization: Assessment of potential of medical devices to produce delayed contact dermatitis Methods of test for skin irritation of extracts from medical devices Method of test for skin irritation of solid medical devices Method of test for dental materials Method of test for eye irritation Method of test for toxicity to cells in culture of extracts from medical devices

Part 2 of this standard provides guidance on the selection of recommended methods of test for initial assessment of biological hazards in medical devices which has been referred to in this standard. In the preparation of this sta-dard, assistance has been derived from BS 5736 : Part 5 : 1982 `Evaluation of medical devices for bIologica hazards: Part 5 Method of test fur systemic toxicity; Assessment of pyrogemcity in rabbits of extracts from medical devices', issued by the British Staudnrds Institution, UK.

IS 12572 ( Part 6 ) : 1988

Indian Standard GUIDE FOR EVALUATION OF MEDICAL DEVICES FOR BIOLOGICAL HAZARDS PART 6 METHOD OF TEST FOR SYSTEMIC TOXICITY : A S S E S S M E N T
OF PYROGENICITY IN RABBITS OF EXTRACTS FROM MEDICAL DEVICES 2.2 Initial Temperature The mean of two temperatures taken at a 30 ~h.JkS interval in the 40 minutes p r e c e d i n g injecting of the extract. 2.3 Maximum Temperature The highest temperature recorded in 3 hours following the injection of the extract. 2.4 Medical Device Any item used in medical treatment, diagnosis or contraception, not intended to have a pharmacoJo_ pical action on the body. 2.5 Pyrogen A fever producing substance. 2.6 Response The difference between the maximum temperature and the initial temperature of each rabbit. 2.7 Summed Response 1 SCOPE 1.1 This standard ( Part 6 ) describes a method of test designed to identify the presence of pyrogenic materials in or on medical devices. This method specifies measurement of the rise in body temperature of rabbits during a period of 3 h following an intravenous injection of an extract of the device, prepared with pyrogen-free saline. 1.2 This method of test is recommended for the initial assessment of devices in Categories A, B1, C and E, ( see Part 2 of this standard ): a) Category A - Those devices intended for long-term ( long-term covers a period from a few months to permanent use ) implantation within the body tissue, for example, artery grafts and hip prostheses; b) Category 4 - Those devices intended for long-term ( long-term covers a period from a few months to permanent use ) or repeated contact with mucosal surfaces, for example, contraceptive devices;

4

Category C - Those devices intcndcd for short-term ( short-term covers a period from a few minutes to several weeks continuous use ) use within the body or in contact with the mucosal surfaces, for example, tracheal tubes, urinary catheters and intravenous cannulae; Category El -- Those devices intended to contain or administer substances, including blood, that are introduced paranterally into the body, for example, blood bags, hemodialysis equipment, intravenous administration sets and kits and hypodermic syringes.

The sum of responses of the rabbits selected for the main test. 2.8 Test Material The final product or sample of final product that is to be tested. 3 ANIMALS AND HUSBANDRY 3.1 p, minimum of three healthy adult rabbits of either sex, weighing not less than 1'5 kg each, shall be used ( SW 5.3.1 ). 3.2 The animals shall be housed individually in au area of uniform temperature, co~ltrollcd tu within +j"C, alld kept free from disturbanccs likly to excite them. 3 . 3 The animals shall IMVC free ~CCCSS to fi)od and water, except as specified in 5.3.2 alld 5.3.3. The diet shall not contain antlbiolicb.

d)

2 DEFINITlONS For the purpose of this standard, the following definitions shall apply. 2.1 Final Product Medical device in its ready-for-use state.

IS 12572 ( Part 6 ) : 1988 4 PREPARATION OF EXTRACTS 4.1 Sterilkation and Handling of Test Material 4.1 .I Pyrogen-free glassware, syringes and needles sh;,ll bc Llsed throughout the test procedure. 4.1.2 Test material that IS, pre-sterilized ( refers to devices obtained from the manufacturer o r sLIpplier sterilized ) shall be handled aseptic::lly throughout the extr`iction and test procedure. 4.1.3 Test materiel that is normally supplied nonsterile but is sterilized bet'ore use shall he sterilized before the extraction procedure and handled aseptically throughout the extraction and lest procedure. 4.1.4 Test material not required to Fc sterile in LISC sII;III either be prepared as specifird in 4.1.3 or if such devices cannot be sterilized, the exlractlon p r o c e d u r e :I:all te corrietl out at r'ct less than ~(J"C to minimize bacterial growth, and Lhe extracts obtained shall be sterilized before administration. 4.2 Extractant Pyrogen-free sterile saline s o l u t i o n c o n t a i n i n g 9 gil sodium chloride shall be used. 4.3 Principles of Extraction 4.3.1 The test material si~all be representative of the iinal product. 4.3.1.1 To achieve this, the test material m a y consist of one or mere intact devices or, If more appropriate, may consist of small pieces of the linal product. For large multicomponcnt CkvJcCS ( for cxamplc, dialysis unit5 or membrane oxlgtnators ), it m:iy be appropriate to separately test the individual components of the device. 4.3.2 If possible, the volume 01` ext~actant selected sho~~ld b e s u c h that it pelni;t3 lr>Jcctlon i n t o a
slll&!le

4.4 Procedure
c a r r y out

4.4.1 Prepare a sufficient volume of extract to the t e s t prccedLlre o n three rabbits ( see 7.5 ).

4.4.2 Ensure a tine of contact between extractant and sample wh:ch, wlthin the requirenients of 4.1.4 simulates the conditions under chicly the dcbicc will normally be used, except that it shall be n o t less than 2 h and need not be lo~;ge~ than 72 h. 4.4.3 Sterilize any extract obtained from unsterilized devices ( set 4.1.4 ). 4.4.4 Record for the test report, the prcciae conditions of extraction. 5 TEST PROCEDL'RE 5.1 Apparatus 5.1 .I R&hit Retuinirlg Boxes
These shall be of open design with loose-fitting

neck stocks so as to permit the animals to sit in A normal resting position.

These s h a l l h a v e a n a c c u r a c y o f I 0'1' C.

5.2 Suitability of Animals A rabbit shall not be used in the pyrogenicit:y test if: a) it has been used in the prcreding three da>s for a p),rogeniciLy test; or b) it has been used, in the prcccding t h r e e weeks, for a pyrogenicit) Lest 111 which the extract under examinativn pruduccd :1 s~Jmrnec1 response cxcecding I ' IS C`: or c) it has been used, at any time, for a pyr(lgcnlcity test in which the lllCill1 I'Cbp_~llbC cl` the rabbits in tile b:itch cxcccci~d 1'2 C`; UI
d) :~ny typical body weight Iohs has occurred

I-abht ol'

n o t

less than 90 pclCLllt of the

extract obtained f?om a sin@ device. 4.3.3 The technique used in exposing the test material to the cxtractant shall bc selcc~~d :.ccw ding to the rcquircmcnts of 4.3.1 and 4.3.2, a n d may consist of total immersion or tlow-through methods, 01. ;I succc'ssive cxtraclion of' mL:rc tllan one spccimcn of the fin:il product. 4.3.4 Extr;!ction conditions, within the rcquircmen& of 4.1.4, sha11 simulate as nearly as possible the conditions unclcr \2hich the device will normally 1-c used. 4.3.5 D u r i n g the cxtrnction, 2 7 ; 3 C shall bc representative of room ternperatLuc and 37-i 2 C that of the body LcmreratuIc.

during the week preceding LIIC that. 5.3 Prcparatiou of Animals

IS 12572 ( Part 6 ) : 1988 5.3.3 Withhold water from the rabbits during the test. 5.3.4 Maintain the rabbits throughout the test procedure, in environmental conditions similar to those in which the animals are housed. 53.5 Weigh the rabbits and record the results. 5.3.6 Not less than one hour before any tcmperature measurement. transfer each rabbit to a retaining box and infert a thermometer into the rectal tract to a depth of between SU and 75 mm. U SC appropriate me;ins to maintain the thcrmomelcr in a fixed position throughout the tests. 5.4 Preliminary Test 5.4.1 Conduct the preliminary test, one to three days befort: the m:~in test. 5.4.2 Warm the pyrogen-free sterile saline solution to 38 5*1%"c. c) A rabbit shall be excluded from the test, I[ the initial tempcruturc is not between 3X.0 C and 39'8°C. 5.5.3 Select three rabbits whose initial tcmpcratures before Injection do not drlrer by more Ihan
1'0 c.

5.5.4 Warm the extract to 38'5 1 1'0°C. 5.5.5 Inject, as described in 5.4.4, the extract into each of the three rabbits. 5.5.6 Record the temperature of each rabbit at 30 minutes intervals for 3 hours after illjcction and note the maximum temperature ( .sw 2.3 ). 6 PRESENTATION OF RESULTS

The results shall be submitted in a test report that includes a complete record of the method of extraction and procedures followed and any other 5.4.3 Record the temperature of each rabbit at 30 relevant data used in the assessment of results as minutes intervals. beginning 90 minutes before described iiijection. 5.4.4 Slowly inject via the marginal ( lateral ) vein 7 ASSESSMENT OF RESULTS of one car of cacti rabbit, 10 ml/kg of pyrogen-free stcrilc saline solution. The duration of injection 7.1 The assessment of the results should be carried out by an expel-ienced person ( preferably a toxishnll not exceed 4 minutes. colo_cist ) who is aware of the conditions of use of 5.4.5 Continue to record the temperature of each the final product and has with him the appropriate rabbit at 3t.l nlinutes intervals for 3 hours after chemical and biological data concerning It ( JW 3.3 injection. and 3.4 of Part 2 ot` this standard ). 5.4.6 Reject from use in the main test all animals which show a dillkrcnce in excess of O'h"C between 7.2 The overall assessment of the pyrogenicity of medical device shall be carried out hy taking into the highest and the lowest rccordcd tcmpcraturcs. consideration all the information giben in the ~cst rcpor t. 5.5 Rain Test
j,, 7,

5.5.1 R e c o r d t h e prc-inicction tcnipcr:itures of each rabbit at 30 mlnutcs iiltervills, brginning not tens than 90 minuks hcfore injection and determine the initial tcnlpcr'atul'c ( .sw 2.2 ). 5.5.2 Identify those rabbits which are to receive the test extract accordin g to the follow~ing criteria: a) A rabhtt sh;~ll he excluded from the test, if the clificrcnce hclwccll winy two conscculivc rcadii~gs exceeds 0'2 C. b) A rabbit sll,~II he cxclu&d from the test, il` the range vf temperature re;idings excccdb 0 4 c.

7.3 Nepativc values for the rcsponsc shall be t:tkcn as zero. 7.4 Assessment shall be based on the ~~~mmcd respunses of thu selcctcd rabbtts.

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__I-----

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